Li Q, Li Y, Xie Y
Research Institute of Tropical Medicine, First Military Medical University, Guangzhou.
Zhonghua Yi Xue Za Zhi. 1995 Jun;75(6):325-8, 381.
A hybrid gene named HGFC coding three protective antigenic epitopes of Plasmodium falciparum and two exogenous T cell activating epitopes was designed and synthesized. A multicopy hybrid gene named HGF-CAC was also constructed. The two genes were cloned into expression vector pWR450-1 and the hybrid fusion proteins containing forgine antigens and beta-galactosidase were expressed in E. Coli. The molecular weights of the fusion proteins were 65KDa and 77KDa respectively. The expression rate was about 35% of total bacterial proteins. The fusion protein could react specifically with mouse and rabbit antibodies against antigens of Plasmodium falciparum. The rabbit immune serum against the purified fusion protein could specifically recognize the antigens of Plasmodium falciparum and effectively inhibit the in vitro development of the parasites. The inhibitory capacity of the immune sera to parasite invasion was enhanced as the amount of the sera increased and the incubation time of the sera with the parasites was prolonged. After 72h incubation at 20% concentration with the parasites, the serum suppressed the multiplication of parasite to a level of 82% and caused degeneration and death of the parasites. The results indicated that the recombinant hybrid antigen of Plasmodium falciparum has immunological activity and protectivity. It is probably a candidate malaria vaccine.
设计并合成了一个名为HGFC的杂种基因,其编码恶性疟原虫的三个保护性抗原表位和两个外源性T细胞激活表位。还构建了一个多拷贝杂种基因HGF-CAC。将这两个基因克隆到表达载体pWR450-1中,含外源抗原和β-半乳糖苷酶的杂种融合蛋白在大肠杆菌中得以表达。融合蛋白的分子量分别为65KDa和77KDa。表达量约占细菌总蛋白的35%。该融合蛋白能与小鼠和兔抗恶性疟原虫抗原的抗体发生特异性反应。兔抗纯化融合蛋白免疫血清能特异性识别恶性疟原虫抗原,并有效抑制疟原虫的体外发育。免疫血清对疟原虫入侵的抑制能力随血清量的增加和血清与疟原虫孵育时间的延长而增强。在20%浓度下与疟原虫孵育72小时后,血清将疟原虫增殖抑制到82%的水平,并导致疟原虫变性和死亡。结果表明,恶性疟原虫重组杂种抗原有免疫活性和保护作用。它可能是一种候选疟疾疫苗。